工程化氨酸氧化酶与有效的辅因子合成和再生系统相结合,用于高产量的5-基三丰的生产
Yulin Ai1, Yusong Huang2, Hongru Zhao2
1College of Biological Science and Engineering, Fuzhou University, Fuzhou, 350108, China.
Bioresources and bioprocessing
|March 6, 2025
概括
使用工程化大肠杆菌,生物衍生5-基酸 (5-HTP) 的产量得到了显著的改善. 这项研究实现了通过发酵实现工业5-HTP生物合成的创纪录的13.9 g/L产量.
科学领域:
- 生物技术和代谢工程 生物技术和代谢工程
- 微生物发酵 微生物发酵
- 酵素工程是什么意思 酵素工程
背景情况:
- 5-基 (5-HTP) 是一种治疗睡眠障碍的天然药物,生物来源是安全的首选.
- 目前的微生物生产方法产生低位数 (<10 g/L),阻碍了工业化.
- 有效的5-HTP生物合成需要强大的微生物菌株和优化的酶转化.
研究的目的:
- 开发一种工程化的大肠杆菌菌株,用于高滴度5-HTP生物合成.
- 通过蛋白质工程优化L-Tryptophan到5-HTP的酶转化.
- 通过代谢工程和发酵过程优化来增强5-HTP的生产.
主要方法:
- 通过删除降解和分支途径和抑制系统,构建了一个积累托的大肠杆菌菌株 (TRP1).
- 使用氨酸氧化酶突变体 (XcPAH) 和MH4再生系统选并设计了一种氧化模块.
- 采用基因组编辑来增强产品流量和辅助因子供应,随后进行料批发发酵.
主要成果:
- 工程菌株TRP1-XC4在基酶的蛋白质工程后产生了319.4mg/L的5-HTP.
- 最后的工程菌株TRP5-XC4在料批发发酵中实现了创纪录的13.9g/L 5-HTP的产量.
- 实现了0.29g/L/h的时空产量,代表了5HTP生物合成中报告的最高产量.
结论:
- 成功开发了一种工程化大肠杆菌菌株,用于高效的5-HTP生物合成.
- 建立了一种绿色和工业上可行的生产5-HTP的方法.
- 取得的产量和生产率为大规模的5-HTP生物生产铺平了道路.
更多相关视频
09:31PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase
Published on: September 26, 2020
4.2K
08:33Radiosynthesis of 1-2-[18F]Fluoroethyl-L-Tryptophan using a One-pot, Two-step Protocol
Published on: September 21, 2021
3.0K
相关概念视频
Amino Acid Biosynthetic Pathways
Amino acid biosynthesis is essential for cell growth, protein synthesis, and metabolic regulation. Cells generate essential and non-essential amino acids from metabolic intermediates to sustain vital biological functions. These intermediates originate from key metabolic pathways: glycolysis, the tricarboxylic acid (TCA) cycle, and the pentose phosphate pathway. Important precursors include α-ketoglutarate, pyruvate, oxaloacetate, phosphoenolpyruvate, and erythrose-4-phosphate, which provide...
Repressible Operon: trp Operon
The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...
Production of Antibiotics
Penicillin, one of the earliest and most widely used antibiotics, is produced industrially by the filamentous fungus Penicillium chrysogenum. Large stirred-tank bioreactors ranging from tens to hundreds of thousands of liters maintain tightly controlled temperature, pH, and dissolved oxygen conditions to support fungal metabolism and maximize antibiotic yield. Penicillin is a secondary metabolite, synthesized primarily during the stationary growth phase, which requires a carefully managed...
Production of Pharmaceuticals
Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...
